ZFIN ID: ZDB-FIG-150427-55
Liu et al., 2015 - Evolutionarily conserved regulation of hypocretin neuron specification by Lhx9. Development (Cambridge, England)   142(6):1113-24 Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Fish:
Knockdown Reagents:
Anatomical Term:
Stage: Prim-5
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Prim-5

Fig. 6

Lhx9 is required for Hcrt neuron specification. (A-C) hcrt ISH at 24hpf shows that morpholino-mediated knockdown of lhx9 reduces the number of Hcrt neurons and the level of hcrt expression (A) compared with embryos injected with a control morpholino (B). (C) Quantification of Hcrt neurons per brain hemisphere at 24, 48 and 72hpf revealed that lhx9 morphants have ~40% fewer Hcrt neurons. Co-injecting a p53 morpholino did not affect this phenotype. Mean±s.e.m. is shown. At least 22 embryos were quantified for each condition. ***P<0.001 compared with control morpholino (one-way ANOVA followed by Bonferroni′s correction for multiple comparisons). (D-G) hcrt ISH at 24hpf shows that co-injection of Cas9 protein and ten lhx9 sgRNAs eliminates hcrt expression (D) compared with embryos injected with Cas9 alone (E). (F) Quantification of Hcrt neurons per brain hemisphere at 24hpf. Mean±s.e.m. is shown. ***P<0.001 compared with Cas9 alone (one-way ANOVA). (G) The percentage of brain hemispheres containing the indicated number of Hcrt neurons. n, number of brain hemispheres analyzed in F,G. Arrowheads indicate endogenous Hcrt neuron region. Scale bars: 50µm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
hcrt WT control Prim-5 hypocretin-secreting neuron ISH
WT + CRISPR1-lhx9 + CRISPR10-lhx9 + CRISPR2-lhx9 + CRISPR3-lhx9 + CRISPR4-lhx9 + CRISPR5-lhx9 + CRISPR6-lhx9 + CRISPR7-lhx9 + CRISPR8-lhx9 + CRISPR9-lhx9 standard conditions Prim-5 hypocretin-secreting neuron ISH
WT + MO2-lhx9 standard conditions Prim-5 hypocretin-secreting neuron ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + CRISPR1-lhx9 + CRISPR10-lhx9 + CRISPR2-lhx9 + CRISPR3-lhx9 + CRISPR4-lhx9 + CRISPR5-lhx9 + CRISPR6-lhx9 + CRISPR7-lhx9 + CRISPR8-lhx9 + CRISPR9-lhx9 standard conditions Prim-5 hypocretin-secreting neuron decreased amount, abnormal
WT + MO2-lhx9 standard conditions Prim-5 hypocretin-secreting neuron decreased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Development (Cambridge, England) for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Development